The expression of CD40 and CD40L on the surface of peripheral blood mononuclear cells in asthmatic rats and the effect of anti-CD40L McAb on Th1 and Th2 cytokines

Objective： To investigate the expression of CD40 and CD40 ligand （CD4OL） on the surface of peripheral blood mononuclear cells（PBMCs） in asthmatic rats and the effect of anti-CD40L McAb on cytokines of PBMCs. Methods： Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs in asthmatic rats. After the PBMCs was treated with anti-CD40L McAb, ELISA was used to detect the IL-4 and IFN-γ levels of culture supernatants. Results： Compared with the normal control group, the expression of CD40 and CD40L of PBMCs in asthmatic rats increased （P 〈 0.05）. Compared with the untreated group, the level of IL-γ and the ratio of IL-4/IFN-γ decreased after the PBMCs was treated with anti-CD40L McAb（P 〈 0.05）. Conclusion： The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats was up-regulated. Anti-CD40L McAb can rectify the imbalance of Th1 and Th2 cytokines.

Experimental Studyon Anti－tumor Effect of SplenocytesInduced by Anti－CD3McAb，PHA and IL－2

The proliferation of splenocytes from healthy adults was induced by anti-CD3 McAb,PHA and IL-2.The proliferative capability and anti-tumor activity as well as phenotypes of the splenocytes cultured in different medium systems were studied.The results showed that anti-CD3 McAb and PHA not only enhanced the proliferation of splenocytes induced by IL-2,but also produced synergism if used simultaneously.The expressions of CD4 and Tac of cellular surface markers were increased after splenocytes were induced by anti-CD3 McAb and PHA.The results of anti-tumor activity of LAK cells suggested that PHA had the capability to promote anti-tumor activity of LAK cells by both direct and in direct pathways,but anti-CD3 McAb indirectly promoted anti-tumor activity of LAK cellsby enhanctng splenocyte proliferation.

抗CD28+B7.1单抗共刺激T细胞诱导肝癌细胞凋亡的第二信使系统研究

目的 :探讨T淋巴细胞活化、增殖、诱导肝癌细胞凋亡过程中第二信使系统的调控机制。方法 :用CD2 8+B7 1(CD80 )单克隆抗体共刺激T淋巴细胞诱导肝癌细胞 (BLE 740 2 )凋亡 ,测定不同诱导时间T细胞内cAMP、cGMP和Ca2 + 的浓度改变及肝癌细胞凋亡情况。结果 :活化的T淋巴细胞中 ,cAMP浓度在初期短暂升高 ,继而快速下降 ,作用于肝癌细胞后逐步回升至 1～ 2倍 ,而细胞内cGMP的浓度急剧升高 6～ 7倍 ,Ca2 + 浓度也显著增加 2～ 3倍 ,且均在第 4天达最高值 ,与癌细胞的凋亡呈正相关。结论 :T淋巴细胞内第二信使系统cAMP、cGMP和Ca2 + 的水平与细胞的活化增殖、杀伤效应密切相关。

抗CD28+B7.1单抗共刺激PBLs诱导肝癌细胞凋亡的蛋白激酶信号转导

目的研究 T淋巴细胞活化、增殖、介导肝癌细胞凋亡过程中其蛋白激酶 C(PKC)和酪氨酸蛋白激酶 (TPK)的活性变化。方法用抗 CD2 8+ B7.1(CD80 )单克隆抗体共刺激正常人外周血淋巴细胞 (PBL s)后作用于肝癌细胞 (BEL -740 2 )。结果激活的 T细胞中 PKC、TPK活性明显增强 ,并与肝癌细胞凋亡程度呈正相关。结论 PKC、TPK在 T细胞活化。

免疫因子对慢性免疫性血小板减少性紫癜骨髓巨核祖细胞体外生长的影响

目的 :探讨慢性免疫性血小板减少性紫癜 (CITP)骨髓巨核祖细胞生长成熟与免疫因素的关系。方法 :通过巨核祖细胞体外培养 ,观察CITP骨髓巨核祖细胞生长和成熟情况 ,并观察抗CD80抗体及干扰素α 2b(IFN α 2b)对CITP和对照组巨核祖细胞生长和成熟的影响 ,并与对照组比较。结果 :①CITP组的各种集落数(CFU MK、BFU MK、mCFU MK)均低于对照组 ,两组间集落数的均值比较差异有统计学意义 (P <0 .0 5 )。CITP组的直径和面积均低于对照组 ,两组间直径和面积均值比较差异有统计学意义 (P <0 .0 5 )。②加入抗CD80抗体的CITP的巨核祖细胞集落生成数明显增多 (P <0 .0 5 )。③IFN α 2b对两组的总集落均有抑制 ,CITP比对照组的巨核祖细胞集落生成受抑制程度低 (P <0 .0 5 )。结论 :CITP巨核祖细胞的集落生成和成熟度较低。抗CD80抗体和IFN α

应用环磷酰胺研制细胞表面抗原的单克隆抗体

利用环磷酰胺和杂交瘤技术 ,成功制备了 1株稳定分泌抗人CD80 分子单克隆抗体的杂交瘤细胞。实验结果表明 ,该单抗腹水效价为 1 0 6 ,属IgG1亚类 ,特异性强 ,为今后的应用打下了基础。